The present invention relates generally to the detection and treatment of Chlamydial infection. In particular, the invention is related to polypeptides comprising a Chlamydia antigen and the use of such polypeptides for the serodiagnosis and treatment of Chlamydial infection.
Chlamydiae are intracellular bacterial pathogens that are responsible for a wide variety of important human and animal infections. Chlamydia trachomatis is one of the most common causes of sexually transmitted diseases and can lead to pelvic inflammatory disease (PID), resulting in tubal obstruction and infertility. Chlamydia trachomatis may also play a role in male infertility. In 1990, the cost of treating PID in the US was estimated to be $4 billion. Trachoma, due to ocular infection with Chlamydia trachomatis, is the leading cause of preventable blindness worldwide. Chlamydia pneumonia is a major cause of acute respiratory tract infections in humans and is also believed to play a role in the pathogenesis of atherosclerosis and, in particular, coronary heart disease. Individuals with a high titer of antibodies to Chlamydia pneumonia have been shown to be at least twice as likely to suffer from coronary heart disease as seronegative individuals. Chlamydial infections thus constitute a significant health problem both in the US and worldwide.
Chlamydial infection is often asymptomatic. For example, by the time a woman seeks medical attention for PID, irreversible damage may have already occurred resulting in infertility. There thus remains a need in the art for improved vaccines and pharmaceutical compositions for the prevention and treatment of Chlamydia infections. The present invention fulfills this need and further provides other related advantages.
The present invention provides compositions and methods for the diagnosis and treatment of Chlamydia infection. In one aspect, polypeptides are provided comprising an immunogenic portion of a Chlamydia antigen, or a variant of such an antigen. In one embodiment, the antigen comprises an amino acid sequence encoded by a DNA sequence selected from the group consisting of (a) a sequence of SEQ ID NO: 1, 15, 21-25, 44-64, 66-76 or 79-88; (b) the complements of said sequences; and (c) sequences that hybridize to a sequence of (a) or (b) under moderately stringent conditions. In a specific embodiment, a polypeptide comprising an amino acid sequence of SEQ ID NO: 5 is provided.
In a related aspect, polynucleotide sequences encoding the above polypeptides, recombinant expression vectors comprising one or more of these polynucleotide sequences and host cells transformed or transfected with such expression vectors are also provided.
In another aspect, the present invention provides fusion proteins comprising an inventive polypeptide, or, alternatively, an inventive polypeptide and a known Chlamydia antigen. In yet another aspect, the present invention provides antibodies, both polyclonal and monoclonal, that bind to the polypeptides described above.
Within other aspects, the present invention provides pharmaceutical compositions that comprise one or more Chlamydia polypeptides disclosed herein, or a polynucleotide molecule encoding such a polypeptide, and a physiologically acceptable carrier. The invention also provides vaccines comprising one or more of the disclosed polypeptides and a non-specific immune response enhancer, together with vaccines comprising one or more polynucleotide sequences encoding such polypeptides and a non-specific immune response enhancer.
In yet another aspect, methods are provided for inducing protective immunity in a patient, comprising administering to a patient an effective amount of one or more of the above pharmaceutical compositions or vaccines.
In yet a further aspect, methods for the treatment of Chlamydia infection in a patient are provided, the methods comprising obtaining peripheral blood mononuclear cells (PBMC) from the patient, incubating the PBMC. with a polypeptide of the present invention (or a polynucleotide that encodes such a polypeptide) to provide incubated T cells and administering the incubated T cells to the patient. The present invention additionally provides methods for the treatment of Chlamydia infection that comprise incubating antigen presenting cells with a polypeptide of the present invention (or a polynucleotide that encodes such a polypeptide) to provide incubated antigen presenting cells and administering the incubated antigen presenting cells to the patient. In certain embodiments, the antigen presenting cells are selected from the group consisting of dendritic cells, macrophages, monocytes, B-cells, and fibroblasts. Compositions for the treatment of Chlamydia infection comprising T cells or antigen presenting cells that have been incubated with a polypeptide or polynucleotide of the present invention are also provided.
In further aspects of the subject invention, methods and diagnostic kits are provided for detecting Chlamydia infection in a patient. In one embodiment, the method comprises: (a) contacting a biological sample with at least one of the polypeptides or fusion proteins disclosed herein; and (b) detecting in the sample the presence of antibodies that bind to the polypeptide or fusion protein, thereby detecting Chlamydia infection in the biological sample. Suitable biological samples include whole blood, sputum, serum, plasma, saliva, cerebrospinal fluid and urine. In one embodiment, the diagnostic kits comprise one or more of the polypeptides or fusion proteins disclosed herein in combination with a detection reagent. In yet another embodiment, the diagnostic kits comprise either a monoclonal antibody or a polyclonal antibody that binds with a polypeptide of the present invention.
The present invention also provides methods for detecting Chlamydia infection comprising: (a) obtaining a biological sample from a patient; (b) contacting the sample with at least two oligonucleotide primers in a polymerase chain reaction, at least one of the oligonucleotide primers being specific for a polynucleotide sequence disclosed herein; and (c) detecting in the sample a polynucleotide sequence that amplifies in the presence of the oligonucleotide primers. In one embodiment, the oligonucleotide primer comprises at least about 10 contiguous nucleotides of a polynucleotide sequence peptide disclosed herein, or of a sequence that hybridizes thereto.
In a further aspect, the present invention provides a method for detecting Chlamydia infection in a patient comprising: (a) obtaining a biological sample from the patient; (b) contacting the sample with an oligonucleotide probe specific for a polynucleotide sequence disclosed herein; and (c) detecting in the sample a polynucleotide sequence that hybridizes to the oligonucleotide probe. In one embodiment, the oligonucleotide probe comprises at least about 15 contiguous nucleotides of a polynucleotide sequence disclosed herein, or a sequence that hybridizes thereto.
These and other aspects of the present invention will become apparent upon reference to the following detailed description. All references disclosed herein are hereby incorporated by reference in their entirety as if each was incorporated individually.
Sequence Identifiers
SEQ ID NO: 1 is the determined DNA sequence for the C. trachomatis clone 1-B1-66.
SEQ ID NO: 2 is the determined DNA sequence for the C. trachomatis clone 4-D7-28.
SEQ ID NO: 3 is the determined DNA sequence for the C. trachomatis clone 3-G3-10.
SEQ ID NO: 4 is the determined DNA sequence for the C. trachomatis clone 10-C10-31.
SEQ ID NO: 5 is the predicted amino acid sequence for 1-B1-66.
SEQ ID NO: 6 is the predicted amino acid sequence for 4-D7-28.
SEQ ID NO: 7 is a first predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 8 is a second predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 9 is a third predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 10 is a fourth predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 11 is a fifth predicted amino acid sequence for 3-G3-10.
SEQ ID NO: 12 is the predicted amino acid sequence for 10-C10-31.
SEQ ID NO: 13 is the amino acid sequence of the synthetic peptide 1-B1-66/48-67.
SEQ ID NO: 14 is the amino acid sequence of the synthetic peptide 1-B1-66/58-77.
SEQ ID NO: 15 is the determined DNA sequence for the C. trachomatis serovar LGV II clone 2C7-8
SEQ ID NO: 16 is the determined DNA sequence for a first putative open reading frame from C. trachomatis serovar D
SEQ ID NO: 17 is the predicted amino acid sequence encoded by the first putative open reading frame from C. trachomatis serovar D
SEQ ID NO: 18 is the amino acid sequence of the synthetic peptide CtC7.8-12
SEQ ID NO: 19 is the amino acid sequence of the synthetic peptide CtC7.8-13
SEQ ID NO: 20 is the predicted amino acid sequence encoded by a second putative open reading from C. trachomatis serovar D
SEQ ID NO: 21 is the determined DNA sequence for clone 4C9-18 from C. trachomatis LGV II
SEQ ID NO: 22 is the determined DNA sequence homologous to Lipoamide Dehydrogenase from C. trachomatis LGV II
SEQ ID NO: 23 is the determined DNA sequence homologous to Hypothetical protein from C. trachomatis LGV II
SEQ ID NO: 24 is the determined DNA sequence homologous to Ubiquinone Mehtyltransferase from C. trachomatis LGV II
SEQ ID NO: 25 is the determined DNA sequence for clone 4C9-18#2 BL21 pLysS from C. trachomatis LGV II
SEQ ID NO: 26 is the predicted amino acid sequence for 4C9-18#2 from C. trachomatis LGV II
SEQ ID NO: 27 is the determined DNA sequence for Cp-SWIB from C. pneumonia strain TWAR
SEQ ID NO: 28 is the predicted amino acid sequence for Cp-SWIB from C. pneumonia strain TWAR
SEQ ID NO: 29 is the determined DNA sequence for Cp-S13 from C. pneumonia strain TWAR
SEQ ID NO: 30 is the predicted amino acid sequence for Cp-S13 from C. pneumonia strain TWAR
SEQ ID NO: 31 is the amino acid sequence for a 10 mer consensus peptide from CtC7.8-12 and CtC7.8-13
SEQ ID NO: 32 is the predicted amino acid sequence for clone 2C7-8 from C. trachomatis LGV II
SEQ ID NO: 33 is the determined DNA sequence of a clone from C. trachomatis serovar D which shows homology to clone 2C7-8
SEQ ID NO: 34 is the predicted amino acid sequence encoded by the sequence of SEQ ID NO: 33
SEQ ID NO: 35 is the DNA sequence for C.p. SWIB Nde (5xe2x80x2 primer) from C. pneumonia 
SEQ ID NO: 36 is the DNA sequence for C.p. SWIB EcoRI (3xe2x80x2 primer) from C. pneumonia 
SEQ ID NO: 37 is the DNA sequence for C.p. S13 Nde (5xe2x80x2 primer) from C. pneumonia 
SEQ ID NO: 38 is the DNA sequence for C.p. S13 EcoRI (3xe2x80x2 primer) from C. pneumonia 
SEQ ID NO: 39 is the amino acid sequence for CtSwib 52-67 peptide from C. trachomatis LGV II
SEQ ID NO: 40 is the amino acid sequence for CpSwib 53-68 peptide from C. pneumonia 
SEQ ID NO: 41 is the amino acid sequence for HuSwib 288-302 peptide from Human SWI domain
SEQ ID NO: 42 is the amino acid sequence for CtSWI-T 822-837 peptide from the topoisomerase-SWIB fusion of C. trachomatis 
SEQ ID NO: 43 is the amino acid sequence for CpSWI-T 828-842 peptide from the topoisomerase-SWIB fusion of C. pneumonia 
SEQ ID NO: 44 is a first determined DNA sequence for the C. trachomatis LGV II clone 19783.3,jen.seq(1 greater than 509)CTL2#11-3xe2x80x2, representing the 3xe2x80x2 end.
SEQ ID NO: 45 is a second determined DNA sequence for the C. trachomatis LGV II clone 19783.4,jen.seq(1 greater than 481)CTL2#11-5xe2x80x2, representing the 5xe2x80x2 end.
SEQ ID NO: 46 is the determined DNA sequence for the C. trachomatis LGV II clone 19784CTL2xe2x80x9412consensus.seq(1 greater than 427)CTL2#12.
SEQ ID NO: 47 is the determined DNA sequence for the C. trachomatis LGV II clone 19785.4,jen.seq(1 greater than 600)CTL2#16-5xe2x80x2, representing the 5xe2x80x2 end.
SEQ ID NO: 48 is a first determined DNA sequence for the C. trachomatis LGV II clone 19786.3,jen.seq(1 greater than 600)CTL2#18-3xe2x80x2, representing the 3xe2x80x2 end.
SEQ ID NO: 49 is a second determined DNA sequence for the C. trachomatis LGV II clone 19786.4,jen.seq(1 greater than 600)CTL2#18-5xe2x80x2, representing the 5xe2x80x2 end.
SEQ ID NO: 50 is the determined DNA sequence for the C. trachomatis LGV II clone 19788CTL2xe2x80x9421consensus.seq(1 greater than 406)CTL2#21.
SEQ ID NO: 51 is the determined DNA sequence for the C. trachomatis LGV II clone 19790CTL2xe2x80x9423consensus.seq(1 greater than 602)CTL2#23.
SEQ ID NO: 52 is the determined DNA sequence for the C. trachomatis LGV II clone 19791CTL2xe2x80x9424consensus.seq(1 greater than 145)CTL2#24.
SEQ ID NO: 53 is the determined DNA sequence for the C. trachomatis LGV II clone CTL2#4.
SEQ ID NO: 54 is the determined DNA sequence for the C. trachomatis LGV II clone CTL2#8b.
SEQ ID NO: 55 is the determined DNA sequence for the C. trachomatis LGV II clone 15-G1-89, sharing homology to the lipoamide dehydrogenase gene CT557.
SEQ ID NO: 56 is the determined DNA sequence for the C. trachomatis LGV II clone 14-H1-4, sharing homology to the thiol specific antioxidant gene CT603.
SEQ ID NO: 57 is the determined DNA sequence for the C. trachomatis LGV II clone 12-G3-83, sharing homology to the hypothetical protein CT622.
SEQ ID NO: 58 is the determined DNA sequence for the C. trachomatis LGV II clone 12-B3-95, sharing homology to the lipoamide dehydrogenase gene CT557.
SEQ ID NO: 59 is the determined DNA sequence for the C. trachomatis LGV II clone 11-H4-28, sharing homology to the dnaK gene CT396.
SEQ ID NO: 60 is the determined DNA sequence for the C. trachomatis LGV II clone 11-H3-68, sharing partial homology to the PGP6-D virulence protein and L1 ribosomal gene CT318.
SEQ ID NO: 61 is the determined DNA sequence for the C. trachomatis LGV II clone 11-G1-34, sharing partial homology to the malate dehydrogenase gene CT376 and to the glycogen hydrolase gene CT042.
SEQ ID NO: 62 is the determined DNA sequence for the C. trachomatis LGV II clone 11-G10-46, sharing homology to the hypothetical protein CT610.
SEQ ID NO: 63 is the determined DNA sequence for the C. trachomatis LGV II clone 11-C12-91, sharing homology to the OMP2 gene CT443.
SEQ ID NO: 64 is the determined DNA sequence for the C. trachomatis LGV II clone 11-A3-93, sharing homology to the HAD superfamily gene CT103.
SEQ ID NO: 65 is the determined amino acid sequence for the C. trachomatis LGV II clone 14-H1-4, sharing homology to the thiol specific antioxidant gene CT603.
SEQ ID NO: 66 is the determined DNA sequence for the C. trachomatis LGV II clone CtL2#9.
SEQ ID NO: 67 is the determined DNA sequence for the C. trachomatis LGV II clone CtL2#7.
SEQ ID NO: 68 is the determined DNA sequence for the C. trachomatis LGV II clone CtL2#6.
SEQ ID NO: 69 is the determined DNA sequence for the C. trachomatis LGV II clone CtL2#5.
SEQ ID NO: 70 is the determined DNA sequence for the C. trachomatis LGV II clone CtL2#2.
SEQ ID NO: 71 is the determined DNA sequence for the C. trachomatis LGV II clone CtL2#1.
SEQ ID NO: 72 is a first determined DNA sequence for the C. trachomatis LGV II clone 23509.2CtL2#3-5xe2x80x2, representing the 5xe2x80x2 end.
SEQ ID NO: 73 is a second determined DNA sequence for the C. trachomatis LGV II clone 23509.1CtL2#3-3xe2x80x2, representing the 3xe2x80x2 end.
SEQ ID NO: 74 is a first determined DNA sequence for the C. trachomatis LGV II clone 22121.2CtL2#10-5xe2x80x2, representing the 5xe2x80x2 end.
SEQ ID NO: 75 is a second determined DNA sequence for the C. trachomatis LGV II clone 22121.1CtL2#10-3xe2x80x2, representing the 3xe2x80x2 end.
SEQ ID NO: 76 is the determined DNA sequence for the C. trachomatis LGV II clone 19787.6CtL2#19-5xe2x80x2, representing the 5xe2x80x2 end.
SEQ ID NO: 77 is the determined DNA sequence for the C. pneumoniae LGV II clone CpS13-His.
SEQ ID NO: 78 is the determined DNA sequence for the C. pneumoniae LGV II clone Cp_SWIB-His.
SEQ ID NO: 79 is the determined DNA sequence for the C. trachomatis LGV II clone 23-G7-68, sharing partial homology to the L11, L10 and L1 ribosomal protein.
SEQ ID NO: 80 is the determined DNA sequence for the C. trachomatis LGV II clone 22-F8-91, sharing homology to the pmpC gene.
SEQ ID NO: 81 is the determined DNA sequence for the C. trachomatis LGV II clone 21-E8-95, sharing homology to the CT610-CT613 genes.
SEQ ID NO: 82 is the determined DNA sequence for the C. trachomatis LGV II clone 19-F12-57, sharing homology to the CT858 and recA genes.
SEQ ID NO: 83 is the determined DNA sequence for the C. trachomatis LGV II clone 19-F12-53, sharing homology to the CT445 gene encoding glutamyl tRNA synthetase.
SEQ ID NO: 84 is the determined DNA sequence for the C. trachomatis LGV II clone 19-A5-54, sharing homology to the cryptic plasmid gene.
SEQ ID NO: 85 is the determined DNA sequence for the C. trachomatis LGV II clone 17-E11-72, sharing partial homology to the OppCxe2x80x942 and pmpD genes.
SEQ ID NO: 86 is the determined DNA sequence for the C. trachomatis LGV II clone 17-C1-77, sharing partial homology to the CT857 and CT858 open reading frames.
SEQ ID NO: 87 is the determined DNA sequence for the C. trachomatis LGV II clone 15-H2-76, sharing partial homology to the pmpD and SycE genes, and to the CT089 ORF.
SEQ ID NO: 88 is the determined DNA sequence for the C. trachomatis LGV II clone 15-A3-26, sharing homology to the CT858 ORF.
SEQ ID NO: 89 is the determined amino acid sequence for the C. pneumoniae clone Cp_SWIB-His.
SEQ ID NO: 90 is the determined amino acid sequence for the C. trachomatis LGV II clone CtL2_LPDA_FL.
SEQ ID NO: 91 is the determined amino acid sequence for the C. pnuemoniae clone CpS13-His.
SEQ ID NO: 92 is the determined amino acid sequence for the C. trachomatis LGV II clone CtL2_TSA_FL.
SEQ ID NO: 93 is the amino acid sequence for Ct-Swib 43-61 peptide from C. trachomatis LGV II.
SEQ ID NO: 94 is the amino acid sequence for Ct-Swib 48-67 peptide from C. trachomatis LGV II.
SEQ ID NO: 95 is the amino acid sequence for Ct-Swib 52-71 peptide from C. trachomatis LGV II.
SEQ ID NO: 96 is the amino acid sequence for Ct-Swib 58-77 peptide from C. trachomatis LGV II.
SEQ ID NO: 97 is the amino acid sequence for Ct-Swib 63-82 peptide from C. trachomatis LGV II.
SEQ ID NO: 98 is the amino acid sequence for Ct-Swib 51-66 peptide from C. trachomatis LGV II.
SEQ ID NO: 99 is the amino acid sequence for Cp-Swib 52-67 peptide from C. pneumonia. 
SEQ ID NO: 100 is the amino acid sequence for Cp-Swib 37-51 peptide from C. pneumonia. 
SEQ ID NO: 101 is the amino acid sequence for Cp-Swib 32-51 peptide from C. pneumonia. 
SEQ ID NO: 102 is the amino acid sequence for Cp-Swib 37-56 peptide from C. pneumonia. 
SEQ ID NO: 103 is the amino acid sequence for Ct-Swib 36-50 peptide from C. trachomatis. 
SEQ ID NO: 104 is the amino acid sequence for Ct-S13 46-65 peptide from C. trachomatis. 
SEQ ID NO: 105 is the amino acid sequence for Ct-S13 60-80 peptide from C. trachomatis. 
SEQ ID NO: 106 is the amino acid sequence for Ct-S13 1-20 peptide from C. trachomatis. 
SEQ ID NO: 107 is the amino acid sequence for Ct-S13 46-65 peptide from C. trachomatis. 
SEQ ID NO: 108 is the amino acid sequence for Ct-S13 56♯75 peptide from C. trachomatis. 
SEQ ID NO: 109 is the amino acid sequence for Cp-S13 56-75 peptide from C. pneumoniae.